TY - JOUR
T1 - Shotgun sequencing of the human transcriptome with ORF expressed sequence tags
AU - Neto, Emmanuel Dias
AU - Correa, Ricardo Garcia
AU - Verjovski-Almeida, Sergio
AU - Briones, Marcelo R.S.
AU - Nagai, Maria Aparecida
AU - Da Silva, Wilson
AU - Zago, Marco Antonio
AU - Bordin, Silvana
AU - Costa, Fernando Ferreira
AU - Goldman, Gustavo Henrique
AU - Carvalho, Alex F.
AU - Matsukuma, Adriana
AU - Baia, Gilson S.
AU - Simpson, David H.
AU - Brunstein, Adriana
AU - De Oliveira, Paulo S.L.
AU - Bucher, Philipp
AU - Jongeneel, C. Victor
AU - O'Hare, Michael J.
AU - Soares, Fernando
AU - Brentani, Ricardo R.
AU - Reis, Luis F.L.
AU - De Souza, Sandro J.
AU - Simpson, Andrew J.G.
PY - 2000/3/28
Y1 - 2000/3/28
N2 - Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. Systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome.
AB - Theoretical considerations predict that amplification of expressed gene transcripts by reverse transcription-PCR using arbitrarily chosen primers will result in the preferential amplification of the central portion of the transcript. Systematic, high-throughput sequencing of such products would result in an expressed sequence tag (EST) database consisting of central, generally coding regions of expressed genes. Such a database would add significant value to existing public EST databases, which consist mostly of sequences derived from the extremities of cDNAs, and facilitate the construction of contigs of transcript sequences. We tested our predictions, creating a database of 10,000 sequences from human breast tumors. The data confirmed the central distribution of the sequences, the significant normalization of the sequence population, the frequent extension of contigs composed of existing human ESTs, and the identification of a series of potentially important homologues of known genes. This approach should make a significant contribution to the early identification of important human genes, the deciphering of the draft human genome sequence currently being compiled, and the shotgun sequencing of the human transcriptome.
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U2 - 10.1073/pnas.97.7.3491
DO - 10.1073/pnas.97.7.3491
M3 - Article
C2 - 10737800
AN - SCOPUS:12944262414
SN - 0027-8424
VL - 97
SP - 3491
EP - 3496
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 7
ER -